Skip to Main Content
 

Global Search Box

 
 
 
 

Files

File List


RaghavendraYadavalli_Dissertation.pdf (18.09 MB)

ETD Abstract Container

Abstract Header

TRAFFICKING AND BIOCHEMICAL CHARACTERIZATION OF PLASMODIUM FALCIPARUM MAURER'S CLEFT TWO TRANSMEMBRANE PROTEIN

Yadavalli, Raghavendra
http://rave.ohiolink.edu/etdc/view?acc_num=csu1535551647632924

Abstract Details

2018, Doctor of Philosophy in Regulatory Biology, Cleveland State University, College of Sciences and Health Professions.
Plasmodium falciparum virulence proteins and molecules required for assembly of the knobs used for cytoadherence are exported through the Maurer’s clefts which are formed in the erythrocyte cytoplasm immediately following merozoite invasion into the erythrocyte host cell. Proteins participating in knob formation, cytoadherence or immune evasion include variant-surface antigens (VSAs) that are trafficked alone or chaperoned by other parasite proteins through the Maurer’s clefts. In this study, recombinant PfMC-2TM was expressed using HeLa based in vitro human cell free expression system. Further, the stage specific expression, trafficking, solubility and topology of endogenous PfMC-2TM in the parasite and erythrocyte cytoplasm was investigated in 3D7 and FC-27 strains of P. falciparum. Detailed cell biological and biochemical characterizations were performed to identify the mode of PfMC-2TM export from the parasite to erythrocyte cytoplasm. Following Brefeldin A (BFA) treatment of parasites, PfMC-2TM traffic was evaluated using immunofluorescence and western blotting with antibodies reactive with PfMC-2TM. Our data shows that PfMC-2TM is sensitive to BFA treatment in the ring and trophozoite stage. Permeabilization of infected erythrocytes with streptolysin O (SLO) and saponin, showed that the N and C-termini of PfMC-2TM are exposed to the erythrocyte cytoplasm with the central portion of the protein protected in the MC membranes. PfMC-2TM was expressed as early as 4 h post invasion (hpi), was tightly colocalized with Maurer’s cleft resident protein REX-1 and trafficked to the erythrocyte membrane without a change in solubility. PfMC-2TM associates with the infected erythrocyte membrane as an integral membrane protein and is seen to be transiently exposed on the infected red blood cell surface. PfMC-2TM remains insoluble upon association with the MC and erythrocyte membrane, suggestive of protein-lipid interactions with membranes of the MC and erythrocyte. PfMC-2TM is an additional marker of the nascent MCs, that can be used as a clinical marker of early blood stage infection.
Tobili Sam-Yellowe (Advisor)
Girish Shukla (Committee Member)
Roman Kondratov (Committee Member)
Bin Zhang (Committee Member)
Severson Aaron (Other)
Ronald Blanton (Other)
221 p.
Biochemistry; Biology; Parasitology
Plasmodium falciparum; Maurers Clefts; PfMC-2TM; Invitro human cell free expression system; Protein purification

Recommended Citations

Citations

  • Yadavalli, R. (2018). TRAFFICKING AND BIOCHEMICAL CHARACTERIZATION OF PLASMODIUM FALCIPARUM MAURER'S CLEFT TWO TRANSMEMBRANE PROTEIN [Doctoral dissertation, Cleveland State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=csu1535551647632924

    APA Style (7th edition)

  • Yadavalli, Raghavendra. TRAFFICKING AND BIOCHEMICAL CHARACTERIZATION OF PLASMODIUM FALCIPARUM MAURER'S CLEFT TWO TRANSMEMBRANE PROTEIN. 2018. Cleveland State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=csu1535551647632924.

    MLA Style (8th edition)

  • Yadavalli, Raghavendra. "TRAFFICKING AND BIOCHEMICAL CHARACTERIZATION OF PLASMODIUM FALCIPARUM MAURER'S CLEFT TWO TRANSMEMBRANE PROTEIN." Doctoral dissertation, Cleveland State University, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=csu1535551647632924

    Chicago Manual of Style (17th edition)

csu1535551647632924
149
© 2018, all rights reserved.
This open access ETD is published by Cleveland State University and OhioLINK.